Radioactive labeling of recombinant antibody fragments by phosphorylation using human casein kinase II and [gamma-32P]-ATP

Nature Biotechnology
D NeriA M Creighton

Abstract

A wide range of antibody fragments can be expressed in bacteria and detected immunochemically via peptide tags. Using specially designed tags, we have developed a strategy for radiolabeling antibody fragments secreted from bacteria. Tagged antibody fragments were secreted either into the bacterial periplasm or the culture medium. The tag was not subject to proteolysis either in the broth or in human plasma. After affinity purification the antibody fragments were phosphorylated with [gamma-32P]ATP and casein kinase II. The labeled fragments were used in a gel band-shift assay to measure antigen binding affinities. In contrast to non site-specific methods such as radioiodination, antibodies labeled with casein kinase II retain full immunoreactivity. Radioactively phosphorylated antibody fragments may have many other applications, including radioimmunoassays and radioimmunotherapy.

References

Jul 1, 1992·Trends in Biochemical Sciences·D W Carr, J D Scott
Oct 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·J R SawyerF R Blattner
Aug 15, 1991·Nature·T ClacksonG Winter
May 23, 1991·European Journal of Biochemistry·N GrankowskiO G Issinger
Apr 1, 1991·Nuclear Medicine Communications·K E BrittonM Granowska
Feb 13, 1990·Biochemistry·R GlockshuberA Plückthun
Sep 24, 1990·Biochimica Et Biophysica Acta·L A Pinna
May 1, 1989·Analytical Biochemistry·X X ZhaoS Pestka
Jan 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·B L LiS Pestka
Jul 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·K TakioE G Krebs
Dec 13, 1984·Nature·M S NeubergerR O Fox
Feb 24, 1995·Journal of Molecular Biology·D NeriG Winter
May 1, 1993·Proceedings of the National Academy of Sciences of the United States of America·L N Kam-MorganJ F Kirsch
May 27, 1994·Journal of Molecular Biology·M FiginiA D Griffiths
Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·P HolligerG Winter
Apr 1, 1995·Bio/technology·D NeriG Winter

❮ Previous
Next ❯

Citations

Nov 4, 1996·International Journal of Cancer. Journal International Du Cancer·B CarnemollaL Zardi
Nov 20, 1997·Clinical Radiology·K E Britton, M Granowska
Jun 26, 2003·Journal of Molecular Biology·Marcello DoniniEugenio Benvenuto
Sep 1, 2000·Advanced Drug Delivery Reviews·F NilssonD Neri
Aug 1, 1997·Current Opinion in Biotechnology·P Carter, A M Merchant
Jan 31, 2007·Arthritis Research & Therapy·Eveline TrachselDario Neri
May 23, 1998·Experimental Cell Research·L BorsiL Zardi
Jul 11, 2006·International Journal of Cancer. Journal International Du Cancer·Verena GafnerDario Neri
Jul 9, 1999·Journal of Molecular Recognition : JMR·A HuberD Neri
Jun 26, 2007·Rapid Communications in Mass Spectrometry : RCM·Domenico CiavardelliAndrea Urbani
Apr 1, 2005·International Journal of Cancer. Journal International Du Cancer·Christina EbbinghausDario Neri
Oct 12, 2005·International Journal of Cancer. Journal International Du Cancer·Monica FabbriniPaolo Neri
Aug 1, 1995·Cell Biophysics·D NeriG Roncucci
Nov 13, 2001·The British Journal of Dermatology·N YazawaK Tamaki
Dec 1, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·C AvignoloS Parodi
Jan 29, 2016·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Xiaonan JiXiangdong Gao
Jun 15, 1997·Analytical Chemistry·D J AndersonK A Davis
Mar 4, 2008·Molecular Immunology·Maria Elena VillaniMarcello Donini

❮ Previous
Next ❯

Related Concepts

Related Feeds

Bioinformatics in Biomedicine

Bioinformatics in biomedicine incorporates computer science, biology, chemistry, medicine, mathematics and statistics. Discover the latest research on bioinformatics in biomedicine here.